The process of RNA extraction from bacteria, in general, involves an RNA-protective, effective lysis of bacterial cell wall (which may pose difficulties). EDTA promotes loss of outer membrane to provide lysozyme with access to peptidoglycan. Another common method for cell wall lysis is mechanical disruption using a homogenizer (applied for gram-positive bacteria and some strains of gram-negative bacteria). Following lysis, it is necessary to disrupt protein-nucleic acid interactions, which can be achieved by adding sodium dodecyl sulfate (SDS). Next step involves using phenol-chloroform-isoamyl alcohol extraction, where RNA can be obtained from the bottom organic phase, the top phase consists of DNA and the interphase contains proteins. Isoamyl alcohol is an inert and optional addition to this mixture and is added as an anti-foaming reagent to reduce the interphase. Following RNA extraction, the samples should be checked for its quality by gel electrophoresis (23S and 16S rRNAs and 5s rRNA and tRNA bands) or UV spectrophotometric or fluorescence methods.
Get tips on using QuantiFluor® RNA System to perform RNA quantification Fuorimetric - human plasma
Get tips on using miRCURY RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized A549
Get tips on using Absolutely RNA FFPE Kit to perform RNA isolation / purification Tissue - Human Kidney
Get tips on using Total RNA Kit, peqGOLD to perform RNA isolation / purification Tissue - Human Stomach
Get tips on using NucleoSpin® RNA XS to perform RNA isolation / purification Tissue - Human Skin
Get tips on using PAXgene Tissue RNA Kit to perform RNA isolation / purification Tissue - Human Colon
Get tips on using NucleoSpin® RNA/Protein to perform RNA isolation / purification Tissue - Human Cornea
Get tips on using Absolutely RNA Microprep Kit to perform RNA isolation / purification Cells - immortalized EL4
Get tips on using Direct-zol RNA Kits to perform RNA isolation / purification Cells - immortalized VCaP
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment